PROJECT SUMMARY The goal of the NIA Research Career Enhancement Award is to bridge expertise gaps in data science and drug discovery by allowing Alzheimer's disease (AD) researchers to expand their expertise. This application will focus on obtaining both the skillset and preliminary data to transition targets from our innovative approach (The Human Brainome) to mapping risk loci for late onset Alzheimer's disease (LOAD). Rather than looking at a single layer of information as in most genome-wide association (GWAS) studies, our approach involves mapping genomic variation in the context of downstream transcriptomic and proteomic expression. This allows for not only mapping the crucial variation involved in LOAD, but also mapping downstream effects and their directions. Additionally, it allows for building networks of multiple players crucial for disease processes. What is crucial is to obtain models and systems for following the Brainome targets outside of the context of human tissues so that hypothesis that have been generated from the Brainome work can be further tested. These models are crucial for the development of drug screening, as we need manipulatable systems for testing no-go/go outcomes in any de novo drug screen. We proposed to use a novel, innovative approach to iPSC development, by using well-characterized control iPSC lines and both viral overexpression and CRISPR technology to insert pathogenic mutations. The innovation lies in our access to lines from autopsy subjects where pathologically confirmed brain tissue is available for confirmation of central profiles as well as the extensive output data from our screen of the Human Brainome cohorts (AG034504 and AG041232). In Aim 1, we will expand our toolkit of iPSC lines to include 3 additional control lines and 3 late onset Alzheimer's Disease (LOAD) lines. All lines will be stably modified with Cas9, so that CRISPR technologies can be exploited in future work. In Aim 2, the top target from the Brainome screen will be introduced into an additional line from Aim 1 to determine whether our preliminary results replicate. These research aims will be combined with a drug development didactic component with our final project goal of obtaining the necessary skills and experience to apply for further funding to develop these models. This project will be unique from prior iPSC modeling for the following reasons: first, developing lines from postmortem confirmed donors will increase rigor, reproducibility and transparency because background effects and baselines can be determined by chasing back to brain tissue and second, the targets for Aim 2 are innovative since they have never been studied in this context. While to date, the drug development space for LOAD has been rife with failures, given the estimated human health cost of $20 trillion over the next 40 years, it is necessary to continue to pursue treatments. Our targets from the Brainome screen present a unique opportunity to follow outcomes from a hypothesis-free screen in an internationally-based, multiple stage screen of appropriate human brain tissues.